溶酶体在细胞功能和疾病中至关重要，通过酸性鞘磷脂酶 (ASM) 控制其膜完整性，影响癌症进展和治疗耐药性。此外，阳离子两亲性药物（CAD）被称为 ASM 抑制剂并具有抗癌活性，但其与溶酶体膜和 ASM 相互作用的结构机制尚不清楚。我们的研究利用全原子显式溶剂分子动力学模拟，深入研究了糖基化 ASM 与溶酶体膜的相互作用，以及 CAD 代表（即依巴斯汀、羟基依巴斯汀和氯雷他定）对膜和 ASM 的影响。我们的结果证实了 ASM 通过皂苷域与膜的关联，之前仅在粗粒度模型中显示。此外，我们阐明了特定残基和 ASM 诱导的膜曲率在脂质募集和定向中的作用。 CAD 还会在参与膜相互作用的催化域中的环水平上干扰 ASM 与膜的结合。我们的计算方法适用于各种 CAD 或膜成分，可深入了解 ASM 和 CAD 与膜的相互作用，为未来的研究提供有价值的工具。© 2024 由 Elsevier B.V. 代表计算和结构生物技术研究网络发布。

Lysosomes are pivotal in cellular functions and disease, influencing cancer progression and therapy resistance with Acid Sphingomyelinase (ASM) governing their membrane integrity. Moreover, cation amphiphilic drugs (CADs) are known as ASM inhibitors and have anti-cancer activity, but the structural mechanisms of their interactions with the lysosomal membrane and ASM are poorly explored. Our study, leveraging all-atom explicit solvent molecular dynamics simulations, delves into the interaction of glycosylated ASM with the lysosomal membrane and the effects of CAD representatives, i.e., ebastine, hydroxyebastine and loratadine, on the membrane and ASM. Our results confirm the ASM association to the membrane through the saposin domain, previously only shown with coarse-grained models. Furthermore, we elucidated the role of specific residues and ASM-induced membrane curvature in lipid recruitment and orientation. CADs also interfere with the association of ASM with the membrane at the level of a loop in the catalytic domain engaging in membrane interactions. Our computational approach, applicable to various CADs or membrane compositions, provides insights into ASM and CAD interaction with the membrane, offering a valuable tool for future studies.© 2024 Published by Elsevier B.V. on behalf of Research Network of Computational and Structural Biotechnology.